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SNP molecular marker and application for detection of rice resistance to brown planthopper bph14 gene

A technology of molecular markers and brown planthoppers, applied in the field of molecular biology, can solve problems such as environmental pollution, low polymorphism rate, and human hazards, and achieve the effects of environmental friendliness, easy operation, and high sensitivity

Active Publication Date: 2021-02-26
HUAZHI RICE BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main types of markers used in literature reports are SSR and InDel markers, which have the disadvantages of low polymorphism rate and small differences in breeding
The EB or polyacrylamide used in the detection process is easy to pollute the environment and cause harm to the human body

Method used

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  • SNP molecular marker and application for detection of rice resistance to brown planthopper bph14 gene
  • SNP molecular marker and application for detection of rice resistance to brown planthopper bph14 gene
  • SNP molecular marker and application for detection of rice resistance to brown planthopper bph14 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040]Example 1 Development of SNP molecular marker of rice Bph14 gene

[0041]According to the gene location information, the Bph14 gene interval of the brown planthopper resistance gene is located in the interval 35693286-35699010 of rice chromosome 3 (MSU7.0). Take the gene interval as the center and expand 50kb on both sides to extract SNP sites, and select according to the PIC value and whether there are other SNP sites around 50bp of the SNP site. At the same time, according to the cloned Bph14 gene sequence and Nipponbare sequence comparison, the selected SNP site, use BatchPrimer3 to carry out primer design on it. For these SNP markers, 18 rice varieties containing Bph14 gene donor materials RH, IR56 and other Bph14-free rice varieties were tested by KASP reaction, and the SNP marker K_030513 was selected for co-segregation with resistant donor materials and good amplification effect. See the above-mentioned SNP marker development flow chartfigure 1 .

[0042]The primer combinatio...

Embodiment 2

[0059]Example 2 Application of SNP molecular marker of rice Bph14 gene

[0060]According to the molecular marker design of the present invention, the KASP reaction can be used to detect the brown planthopper resistance Bph14 gene in rice materials with high throughput, and the primer combination designed in Example 1 is adopted. Using 95 materials to verify the SNP marker K_030513 in natural populations. The 95 materials included susceptible control materials, common hybrid rice and core rice breeding materials. The results of marking in the natural population classification are as followsfigure 2 As shown, in addition to the heterozygous Bph14 genotype detected by Xieyou 716, other susceptible control materials, common hybrid rice and core rice breeding materials were all detected as non-brown planthopper resistance homozygous Bph14 genotype. It can be seen that SNP marker K_030513 detects the Bph14 gene locus as a highly specific resistance locus, which can be conveniently and effici...

Embodiment 3

[0061]Example 3 The genetic location of SNP markers and the phenotype verification of segregating populations

[0062]1. Mark genetic location verification

[0063]Using 79 Bph14 RIL populations and 11 polymorphic SNP markers in their parents and the Bph14 specific marker K_030513 developed in Example 1 of the present invention for genetic location verification, the marker K_030513 of the present invention was mapped to rice The 79.2cM position of chromosome 3 (seeimage 3 ).

[0064]2. Marked phenotype verification

[0065]The B5 and Mh63 RIL populations were used to verify the genetic phenotype of the marker K_030513 linked to the gene Bph14 in the present invention. The phenotype identification was entrusted to Wuhan Hetaiqing Biotechnology Co., Ltd., 83.3% of the 12 strains containing Bph14 had a phenotype of medium resistance or above, and 83.3% of the 12 strains without Bph14 had a susceptible phenotype. Type (Table 4). The phenotype data corresponds well to the genotype, which again verif...

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Abstract

The invention provides a SNP molecular marker and application for detecting rice brown planthopper resistance gene Bph14. The invention discloses a SNP marker K_030513 with good co-segregation and amplification effect with the rice brown planthopper resistance gene Bph14. The marker detects the base at 35692898 (MSU7.0) of rice chromosome 3, and the polymorphism is C / G, based on KASP The primer sequence of the marker K_030513 developed by the technology is shown in SEQ ID No.1‑3. The SNP molecular marker of the present invention can be used to detect the Bph14 gene locus as a highly specific locus, and can be used conveniently and efficiently to identify whether the rice variety contains the Bph14 gene. The application method of the SNP molecular marker provided by the invention is accurate, reliable, and easy to operate, and is suitable for identification of the Bph14 gene and assisted selection breeding.

Description

Technical field[0001]The invention relates to the technical field of molecular biology, in particular to the SNP molecular marker of the rice brown planthopper resistance gene Bph14 and a method for detecting the molecular marker.Background technique[0002]Rice is an important food crop in our country. The brown planthopper is a monophagous pest of rice. It harms rice plants by sucking the phloem sap through mouth needles, causing slow growth of rice, delayed tillers, and increased empty grains. The brown planthopper is also the transmission vector of some rice viruses such as grassy bushy stunt virus and tooth leaf dwarf virus, which seriously affects the production and safety of rice. At present, the control of brown planthopper mainly relies on chemical control, which not only increases production costs and pest resistance, but also pollutes the environment. Therefore, the use of host resistance to breed brown planthopper-resistant varieties is considered an effective way to contr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 彭佩江南郑秀婷梁毅贺治洲
Owner HUAZHI RICE BIO TECH CO LTD